Catalase: Difference between revisions
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== Application in [[HRR]]Β == | == Application in [[HRR]]Β == | ||
'''Clt: Catalase''' (from bovine liver,lyophilized powder, | '''Clt: Catalase''' (from bovine liver, lyophilized powder; 2,000-5,000 U/mg protein); Sigma C 9322; store at -20 Β°C. | ||
Catalase is included in the mitochondrial respiration medium [[MiPNet14.13_Medium-MiR06 | MiR06]] (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H<sub>2</sub>O<sub>2</sub>. | Catalase is included in the mitochondrial respiration medium [[MiPNet14.13_Medium-MiR06 | MiR06]] (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H<sub>2</sub>O<sub>2</sub>. | ||
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=== Preparation of catalase stock solution === | === Preparation of catalase stock solution === | ||
::'''Stock solution:''' 112 | ::'''Stock solution:''' 112,000 U/ml (dissolved in MiR05) | ||
::'''Example:''' 'Catalase lypophilized powder, | ::'''Example:''' 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein' | ||
::1) Use 'units/mg solid' for your calculations Β | ::1) Use 'units/mg solid' for your calculations Β | ||
::2) Result: 40 mg catalase powder ( | ::2) Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 ml MiR05 to obtain a catalase stock solution with 112,000 U/ml. | ||
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'''Oxygraph-2k manual titrations | '''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::* Titration volume: 5 Β΅l Ctl using a 25 Β΅l syringe | ::* Titration volume: 5 Β΅l Ctl using a 25 Β΅l syringe (2 ml O2k-chamber). | ||
::* Final | ::* Final concentration: 280 U/ml. | ||
Revision as of 11:56, 15 April 2014
- high-resolution terminology - matching measurements at high-resolution
Catalase
Description
Catalase catalyzes the dismutation of hydrogen peroxide to water and oxygen.
Abbreviation: Ctl
Reference: MiPNet09.12
MitoPedia topics: Enzyme
Application in HRR
Clt: Catalase (from bovine liver, lyophilized powder; 2,000-5,000 U/mg protein); Sigma C 9322; store at -20 Β°C.
Catalase is included in the mitochondrial respiration medium MiR06 (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H2O2.
Preparation of catalase stock solution
- Stock solution: 112,000 U/ml (dissolved in MiR05)
- Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'
- 1) Use 'units/mg solid' for your calculations
- 2) Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 ml MiR05 to obtain a catalase stock solution with 112,000 U/ml.
Unit definition: * Units of enzymatic activity (U) in Β΅mol/min; assay used by Sigma Aldrich: ' One unit will decompose 1.0 ΞΌmole of H2O2 per min at pH 7.0 at 25 Β°C, while the H2O2 concentration falls from 10.3 to 9.2 mM, measured by the rate of decrease of A240. '
Oxygraph-2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 5 Β΅l Ctl using a 25 Β΅l syringe (2 ml O2k-chamber).
- Final concentration: 280 U/ml.
